Abstract
Cnidarians are emerging model organisms for cell and molecular biology research. However, successful cell culture development has been challenging due to incomplete tissue dissociation and contamination. In this report, we developed and tested several different methodologies to culture primary cells from all tissues of two species of Cnidaria: Nematostella vectensis and Pocillopora damicornis. In over 170 replicated cell cultures, we demonstrate that physical dissociation was the most successful method for viable and diverse N. vectensis cells while antibiotic-assisted dissociation was most successful for viable and diverse P. damicornis cells. We also demonstrate that a rigorous antibiotic pretreatment results in less initial contamination in cell cultures. Primary cultures of both species averaged 12–13 days of viability, showed proliferation, and maintained high cell diversity including cnidocytes, nematosomes, putative gastrodermal, and epidermal cells. Overall, this work will contribute a needed tool for furthering functional cell biology experiments in Cnidaria.
Highlights
Cnidarians are emerging model organisms for cell and molecular biology research
Primary cell cultures of multiple cell types are important for studying emerging model organisms because they allow for observation and manipulation of a diversity of undescribed cell types that are still functioning as they would in vivo[3,4,5]
We monitored 123 N. vectensis cell cultures and 51 P. damicornis cell cultures. We show in both N. vectensis and P. damicornis that diverse cell types can reliably survive ex vivo for over 12 days
Summary
Cnidarians are emerging model organisms for cell and molecular biology research. successful cell culture development has been challenging due to incomplete tissue dissociation and contamination. Cnidarian tissues are constantly exposed to their natural environment due to relatively simple tissue organization, and this along with their mucus layer, has been hypothesized to lead to a higher association with a diverse microbiome that can overgrow a cell c ulture[16,17,18] Given these challenges there are many areas where cell culture can be improved to achieve longevity. We report the first primary cell cultures using all tissues of the model sea anemone N. vectensis, and the development of a novel method of antibiotic-facilitated dissociation for the adult coral, P. damicornis This is the first time that individual cells from all tissues of coral or sea anemones were shown to survive in cell culture for over 12 days. The goal of this experiment was to build on these previous cnidarian cell culture
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